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241230s2025 xx |||||o 00| ||eng c |
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|a 10.1016/j.plantsci.2024.112376
|2 doi
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|a pubmed25n1391.xml
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|a (DE-627)NLM382198328
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|a (NLM)39733860
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|a (PII)S0168-9452(24)00403-5
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|a DE-627
|b ger
|c DE-627
|e rakwb
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|a eng
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| 100 |
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|a Miao, Wei
|e verfasserin
|4 aut
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| 245 |
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|a A new method for identifying proteins involved in DNA methylation through reverse genetics in Arabidopsis
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|c 2025
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
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|2 rdacarrier
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|a Date Completed 29.04.2025
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|a Date Revised 29.04.2025
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Copyright © 2024. Published by Elsevier B.V.
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|a Forward genetic screens have uncovered numerous genes involved in DNA methylation regulation, but these methods are often time-intensive, costly, and labor-intensive. To address these limitations, this study utilized CRISPR technology to knockout selected co-expressed genes, enabling the rapid identification of low luciferase (LUC) luminescence mutants in the Col-LUC line, which harbors a LUC transgene driven by a 2 × 35S promoter in Arabidopsis. As proof of concept, the repressor of silencing 1 (ROS1) and RNA-directed DNA methylation 1 (RDM1) genes were used as controls, while the increased DNA methylation 3 (IDM3) gene, co-expressed with ROS1, was selected as the target for gene knockout experiments. The results demonstrated that combining co-expression analysis with CRISPR technology is an effective strategy for generating low LUC luminescence mutants in the Col-LUC line. Notably, a new mutant, named reduced luminescence 1 (rl1), was identified through this approach. The rl1 mutant exhibited genome-wide DNA hypermethylation, and its reduced luminescence phenotype was largely reversed by treatment with the DNA methylation inhibitor 5-Aza-2'-deoxycytidine, confirming its anti-silencing role in DNA methylation regulation. This study presents a novel and efficient approach for obtaining low luminescence mutants in the Col-LUC line and identifies RL1 as a previously uncharacterized protein involved in DNA methylation regulation
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|a Journal Article
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|a CRISPR
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|a DNA methylation
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|a Gene co-expression
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|a Reduced luminescence 1 (rl1)
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|a Reverse genetics
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|a Arabidopsis Proteins
|2 NLM
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|a ROS1 protein, Arabidopsis
|2 NLM
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| 650 |
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|a Nuclear Proteins
|2 NLM
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| 700 |
1 |
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|a Dai, Jie
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Zhang, Li
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Liang, Zhile
|e verfasserin
|4 aut
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| 700 |
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|a Sun, Xiaoxuan
|e verfasserin
|4 aut
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| 700 |
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|a Huang, Meizi
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Zhang, Aqin
|e verfasserin
|4 aut
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| 700 |
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|a Zheng, Long
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Li, Yongjun
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Li, Ying
|e verfasserin
|4 aut
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| 773 |
0 |
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|i Enthalten in
|t Plant science : an international journal of experimental plant biology
|d 1985
|g 352(2025) vom: 16. März, Seite 112376
|w (DE-627)NLM098174193
|x 1873-2259
|7 nnas
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| 773 |
1 |
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|g volume:352
|g year:2025
|g day:16
|g month:03
|g pages:112376
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|u http://dx.doi.org/10.1016/j.plantsci.2024.112376
|3 Volltext
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